Tau-Fyn Interaction and Alzheimer's Disease

Tau-Fyn 相互作用与阿尔茨海默病

基本信息

批准号：
10292907
负责人：
Jonathan R Roth
金额：
$ 1.62万
依托单位：
UNIVERSITY OF ALABAMA AT BIRMINGHAM
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-01 至 2021-10-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10292907
关键词：
3-Dimensional AMPA Receptors Ablation Affect Affinity Age Alzheimer&apos s Disease Alzheimer&apos s disease model Alzheimer&apos s disease patient Alzheimer&apos s disease therapeutic Amyloid beta-Protein Automobile Driving Behavioral Assay Binding Biological Assay Biological Models Cognitive Cognitive deficits Dementia Dendrites Dendritic Spines Disease Progression Electroencephalography Epilepsy Functional disorder Genetic Glutamates Goals Hippocampus (Brain)Impaired cognition In Situ In Vitro Knock-out Knockout Mice Lead Learning Ligation MAPT gene Measures Mediating Mediator of activation protein Membrane Modeling Molecular Morphology Mus Mutate Mutation N-Methyl-D-Aspartate Receptors N-Methylaspartate Nerve Degeneration Neurites Neurofibrillary Tangles Neuronal Dysfunction Neurons PXXP Motif Pathogenesis Peptides Phenotype Phosphorylation Phosphotransferases Population Predisposition Proline-Rich Domain Proteins Proto-Oncogene Proteins c-fyn Public Health Role SH3 Domains Scientist Seizures Signal Transduction Structure Synapses System Techniques Testing Therapeutic Vertebral column Work abeta toxicity base behavior test beta amyloid pathology career cognitive testing density excitotoxicity extracellular familial Alzheimer disease gabazine hyperphosphorylated tau immunocytochemistry in vivo inhibitor/antagonist insight member mouse model multi-electrode arrays novel strategies novel therapeutic intervention osmotic minipump overexpression postsynaptic prevent relating to nervous system skills tau Proteins tau interaction therapeutic target therapy development trafficking

项目摘要

Project Summary/Abstract Alzheimer’s disease (AD) is the leading cause of dementia worldwide and its impact will increase exponentially as the population ages. New therapeutic approaches are desperately needed to treat AD. The microtubule- associated protein Tau is has been heavily studies because it aggregates into neurofibrillary tangles within neurons, one of the hallmarks of AD. Genetic knockout of Tau is protective in several models of AD, highlighting its potential as a therapeutic target for AD. Interestingly, Tau reduction also prevents network hyperexcitability, which occurs early in AD and may contribute to neurodegeneration, in these models. Similarly, in a primary neuron culture system, Tau reduction prevents amyloid-β (Aβ) toxicity and glutamate or NMDA-induced excitotoxicity. In short, Tau reduction is protective in a variety of systems, but the mechanism by which it does so is currently unknown. Tau’s central proline-rich region, which is hyperphosphorylated in AD, has several PxxP motifs that mediate binding with SH3 domain–containing proteins including the nonreceptor tyrosine kinase Fyn. Fyn is also an important mediator of network hyperexcitability, as it phosphorylates AMPA and NMDA receptors to strengthen their signaling and regulates dendritic spine dynamics. Exogenous Aβ activates Fyn at the postsynaptic density, leading to NMDAR phosphorylation and excitotoxicity in neurons. Diverse evidence indicates that Tau’s interaction with Fyn could be a critical mediator of Aβ toxicity. Genetic knockout of either Tau or Fyn prevents Aβ toxicity in primary neurons, hyperphosphorylated Tau has a higher affinity for Fyn, and Tau mediates trafficking of Fyn to dendrites in vivo, leading to Aβ-induced cognitive deficits and network hyperexcitability. However, the idea that the Tau-Fyn interaction is a critical mediator of Aβ toxicity has not been directly tested, which is the goal of this project. My overarching hypothesis is that the Tau-Fyn interaction is a critical mediator of Aβ-induced structural and functional abnormalities. I will test this hypothesis using a peptide inhibitor of the Tau-Fyn interaction, Tau-PxxP5/6, developed by previous members of the Roberson lab. I developed a proximity ligation-based assay to confirm that Tau-PxxP5/6 inhibits endogenous Tau-Fyn interaction in situ and found that it prevents Aβ-induced neurite degeneration and membrane trafficking dysfunction. Here, I will determine if Tau-PxxP5/6 prevents Aβ-induced deficits in dendritic spine morphology using 3D morphometric analysis and Aβ- and gabazine-induced network hyperexcitability using multi-electrode arrays. I will also determine if the Fyn-binding region of Tau is the critical region of Tau that mediates Aβ toxicity by transducing Tau knockout neurons with different mutated Tau constructs to prevent Fyn binding. In addition to in vitro studies, I will determine if Tau-PxxP5/6 prevents cognitive deficits, epileptiform activity and seizure susceptibility in vivo using the hAPPJ20 mouse model of AD. The proposed work will provide insights into the molecular mechanisms of Aβ toxicity and could provide a promising therapeutic strategy to treat AD.

项目摘要/摘要阿尔茨海默氏病（AD）是全球痴呆症的主要原因，其影响将成倍增加随着人口的年龄。迫切需要新的治疗方法来治疗广告。微管相关的蛋白质tau是大量研究，因为它聚集在神经纤维缠结中神经元，AD的标志之一。 Tau的遗传敲除在几种AD模型中受到保护，突出显示它作为AD的治疗靶标的潜力。有趣的是，tau降低还可以防止网络过度兴奋性，在AD早期发生，并且可能在这些模型中有助于神经变性。同样，在小学中神经元培养系统，TAU还原可防止淀粉样蛋白β（Aβ）毒性和谷氨酸或NMDA诱导的兴奋性毒性。简而言之，减少tau在多种系统中受到保护，但是它的机制这样做当前未知。 Tau的中央脯氨酸富含AD中的磷酸化的区域具有几个PXXP基序，介导与含Sh3结构域的蛋白质（包括非受体的蛋白）结合酪氨酸激酶Fyn。 FYN也是网络过度兴奋性的重要介体，因为它磷酸化AMPA 和NMDA接收器以增强其信号传导并调节树突状脊柱动力学。外源Aβ 激活突触后密度的FYN，导致神经元中的NMDAR磷酸化和兴奋性毒性。不同的证据表明，陶与FYN的相互作用可能是Aβ毒性的关键介体。遗传 Tau或Fyn的敲除可以预防原发性神经元中Aβ的毒性，高磷酸化的Tau具有较高的对Fyn的亲和力和Tau在体内介导Fyn的贩运，导致Aβ诱导的认知缺陷和网络过度兴奋性。但是，tau-fyn相互作用是Aβ毒性的关键介体的想法尚未直接测试，这是该项目的目标。我的总体假设是tau-fyn 相互作用是Aβ诱导的结构和功能异常的关键介体。我会测试这个使用tau-fyn相互作用的肽抑制剂tau-pxxp5/6的假设，由以前的成员开发罗伯森实验室。我开发了基于连接的近端结扎测定法，以确认tau-pxxp5/6抑制内源性 Tau-fyn的相互作用原位，发现它可以防止Aβ诱导的神经退行性变性和膜运输功能障碍。在这里，我将确定tau-pxxp5/6是否阻止Aβ诱导的树突状脊柱形态缺陷使用3D形态分析以及使用多电极的Aβ-和Gabazine诱导的网络过度兴奋性数组。我还将确定tau的Fyn结合区域是否是介导Aβ毒性的临界区域通过传递具有不同突变tau构建体的TAU基因敲除神经元以防止Fyn结合。此外为了进行体外研究，我将确定tau-pxxp5/6是否可以防止认知缺陷，癫痫样活性和癫痫发作使用AD的HAPPJ20鼠标模型在体内的敏感性。拟议的工作将为您提供见解 Aβ毒性的分子机制，可以提供有希望的治疗AD的治疗策略。